Cell-free expression of functional receptor tyrosine kinases

Wei He; Tiffany M Scharadin; Matthew Saldana; Candice Gellner; Steven Hoang-Phou; Christina Takanishi; Gregory L Hura; John A Tainer; Kermit L Carraway 3rd; Paul T Henderson; Matthew A Coleman

doi:10.1038/srep12896

Cell-free expression of functional receptor tyrosine kinases

Sci Rep. 2015 Aug 14:5:12896. doi: 10.1038/srep12896.

Affiliations

¹ University of California Davis School of Medicine, Radiation Oncology, Sacramento, California, USA.
² University of California Davis School of Medicine, Internal Medicine, Division of Hematology Oncology, Sacramento, California, USA.
³ University of California Davis School of Medicine, Biochemistry and Molecular Medicine, Sacramento, California, USA.
⁴ 1] Lawrence Berkeley National Laboratory, Berkeley, California, USA [2] University of California Santa Cruz, Chemistry and Biochemistry, Santa Cruz, California, USA.
⁵ 1] Lawrence Berkeley National Laboratory, Berkeley, California, USA [2] Scripps Research Institute, La Jolla, California, USA.
⁶ 1] University of California Davis School of Medicine, Biochemistry and Molecular Medicine, Sacramento, California, USA [2] University of California Davis Comprehensive Cancer Center, Sacramento, CA California, USA.
⁷ 1] University of California Davis School of Medicine, Internal Medicine, Division of Hematology Oncology, Sacramento, California, USA [2] University of California Davis Comprehensive Cancer Center, Sacramento, CA California, USA.
⁸ 1] University of California Davis School of Medicine, Radiation Oncology, Sacramento, California, USA [2] University of California Davis Comprehensive Cancer Center, Sacramento, CA California, USA [3] Lawrence Livermore National Laboratory, Livermore, California, USA.

Abstract

Receptor tyrosine kinases (RTKs) play critical roles in physiological and pathological processes, and are important anticancer drug targets. In vitro mechanistic and drug discovery studies of full-length RTKs require protein that is both fully functional and free from contaminating proteins. Here we describe a rapid cell-free and detergent-free co-translation method for producing full-length and functional ERBB2 and EGFR receptor tyrosine kinases supported by water-soluble apolipoprotein A-I based nanolipoprotein particles.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell-Free System
Receptor Protein-Tyrosine Kinases / metabolism*

Substances

Receptor Protein-Tyrosine Kinases

Cell-free expression of functional receptor tyrosine kinases

Authors

Affiliations

Abstract

Publication types

MeSH terms

Substances

Grants and funding